AU641577B2 - Trifluoromethylketone derivatives, processes for preparation thereof and use thereof - Google Patents

Abstract

New trifluoromethylketone derivatives of the following formula : <CHEM> wherein R<1> is lower alkyl which has one or two substituents selected from carboxy, esterified carboxy and di-lower alkylcarbamoyl; phenyl(lower)alkyl, the phenyl moiety of which may have halogen or nitro or amino and the alkyl moiety of which may have carboxy or esterified carboxy; halo-phenyl; morpholino; or morpholino(lower)alkyl, R<2> and R<3> are each lower alkyl, X is - or -NH-, and Y is <CHEM> and pharmaceutically acceptable salt thereof, processes for their preparation and pharmaceutical compositions comprising them.

Description

-7 7 64 577

AUSTRALIA

PATENTS ACT 1990 COMPLETE SPECIFICATION NAME OF APPLICANT(S): Fujisawa Pharmaceutical Co., Ltd.

ADDRESS FOR SERVICE: DAVIES COLLISON CAVE Patent Attorneys 1 Little Collins Street, Melbourne, 3000.

INVENTION TITLE: Trifluoromethylketone derivatives, processes for preparation thereof and use thereof The following statement is a full description of this invention, including the best method of performing it known to me/us:- D ID a' q f) q la- This invention relates to new trifluoromethylketone derivatives.

More particularly, this invention relates to new trifluoromethylketone derivatives and their pharmaceutically acceptable salts which have an human leukocyte elastase-inhibiting activity, to processes for preparation thereof, and to a pharmaceutical composition comprising the same and to a method of use thereof.

"The new trifluoromethylketone derivatives of this invention are represented by the following formula 2 3 R R

(I)

2wherein R 1is lower alkyl which has one or two substituents.

selected from carboxy, esterified carboxy and di-lower alkylcarbanoyl, phenyl(lower)alkyl, the phenyl moiety of which may have halogen or nitro or amino and the alkyl moiety of which may have carboxy or esterified carboxy, halo-phenyl, morph~olino or rorpholino (lower) alkyl,

R

2 and R 3 are each lower alkyl, X is -NH-, n is 0 or 1, and Yis or 11

-NCI,-

According to this invention, the new trifluoromethylketone derivatives and salts thereof can be prepared by various processes which are illustrated by the following reaction schemes Process 1 2. R 3 S. (II)1 too 4 RI-NH 2 (111) *9,09, 2 R 3 :0.

-NHCO -__-ECOHHO CNCcC *5.3 *00S.() 0 0S00 Process 2

R

3 3

OH

3 3 Process 3 R 2

R

3 1 RaNHC04/ [XI ,C~i;u--i.Untuc Sa) 2R 3 R -NHCO-// \-[XnCONHCnuu-x-CuNtHCtCOuF 3 b Process 4 25R2 2 2*

NOHHCC

-H(H04 C) COHCHOCF 2 2 d 3* 4 Process

R

2

R

3 HOOC-R -NHCO-"-[Xn- CONHCHCO-Y-CONICHCOCF 3 (Ie) c 3 (R 2 NH (V)

R

2

R

3 1 0 (R NCO-R-NHCO- E-lXn-

CONHCHCO-Y-CONHCHCOCF

3 (I In the above formulae, R 1 is mono- or di- esterified 1 a carboxy(lower)alky1 and Rb is mono- or dicarboxy(lower)alkyl, R c is lower alkylene, Rd is lower alkyl, and R 1 to R 3 X, Y and n are each as defined above.

A pharmaceuticall'. acceptable salt of the new trifluoromethylketone derivatives of the formula may include a salt with an inorganic or organic base such as an alkali metal salt sodium salt, potassium salt, etc.), an alkaline earth metal salt calcium salt, etc.), ammonium salt, ethanolamine salt, triethylamine 25 addition salt with organic or inorganic acid such as methane sulfonate, hydrochloride, sulfate, nitrate, o phosphate or the like.

ePreferred examples and illustrations of the various *9 30 invention includes within the scope thereof are explained in detail as follows.

atoms, unless otherwise indicated.

Preferred examples of "halogen" is fluorine, T' chlorine, bromine and iodine.

•abx~oe~lyR 5 *0 00 0 00

S@

bose a Preferred examples of "lower alkyl" may include a residue of straight and branched alkane having 1 to 6 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, neopentyl, hexyl and the like, and preferably the one having 1 to 4 carbon atom(s).

Preferred examples of "esterified carboxy" may include an alkyl ester, i.e. alkoxycarbonyl such as lower alkoxycarbonyl methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, etc.) and a phenyl(lower)alkyl ester, i.e.

phenyl(lower)alkoxycarbonyl such as benzyloxycarbonyl and a benzoyl(lower)alkyl ester, i.e.

benzoyl(lower)alkoxycarbonyl such as benzoylmethoxycarbonyl, and the like.

Preferred examples of "lower alkylene" may include methylene, ethylQne, propylene, isopropylene and the like.

Preferred examples of "di-lower alkylcarbamoyl" may include N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl and the like.

Processes for preparing the object compound or its salts of this invention are explained in detail in the following.

In the explanation of Processes 1 to 5 as follows, salts of Compounds (I a to and (II) to may include the same as those exemplified as pharmaceutically acceptable salt of trifluoromethylketone derivatives (I) as illustrated hereinbefore.

e* 0 o* 0 30 Process 1 Compound (II) Compound (III) Compound (I) Compound and its salt can be prepared by reacting Compound (II) or its salt with a Compound (III) or its salt.

6 The reaction of this process can be conducted as follows.

That is, in one case, as the first step, the carboxy group of Compound (II) or its salt is usually activated in a conventional manner, for example, in the form of its acid halide, azide, acid anhydride or a mixed anhydride, activated ester, and the like, and is reacted with the Compound (III) to give Compound and in the other case, the Compound (II) or its salt is reacted with the Compound (III) or its salt directly in the presence of a conventional condensing agent such as N,N-dicyclohe,. .carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and the like.

s

S

0OS S *5 *S S 6* a sees 0* S 0 0*

S

15 This reaction is preferably carried out in a solvent such as N,N-dimethylformamide, methylene chloride, chloroform, tetrahydrofuran, dioxane, ethyl acetate, methanol, ethanol, water or the like under ice-cooling to at ambient tenperature and the reaction in the presence of k condensing agent is usually carried out in an anhydrous, but not critical, conditions.

Process 2 Compound (IV) Compound (I) The Compound and its sa3 can be prepared by oxidizing the Compound (IV) or its salt.

The oxidation is carried out by a conventional method using an oxidizing agent which can be applied to converting a hydroxymethyl group to a carbonyl group such as potassium permanganate, chromic compound chromic acid, sodium chromate, dichromic acid, sodium dichromate, pyridinium chlorochromate, pyridinium dichromate, etc.), Swern reagent (dimethylsulfoxide and oxalylchloride), 7 Jones reagent and the like.

The reaction is usually carried out in a conventional solvent such as water, acetone, dioxane, dimethylformamide, pyridine or any other organic solvents which do not adversely influence to the reaction, or a mixture thereof.

This reaction is preferably carried out under somewhat milder condition such as under cooling, at room temperature or under warming.

Process 3 Compound (Ia) Compound (I b

S

*r 0 00 S 000 50 0 S

S.

sq 00 S 15 The Compound (I b and its salt can be prepared by subjecting the Compound (Ia) or its salt to de-esterification reaction.

The de-esterification reaction is carried out by a conventional method such as hydrolysis, reduction or the like, details of which are explained in the following 0 25 S 30 30 1) Hydrolysis Hydrolysis is preferably carried out in the presence of an acid or base.

Suitable acid includes an inorganic acid (e.g.

hydrochloric acid, hydrobromic acid, sulfuric acid, etc.), an organic acid formic acid, acetic acid, trifluoroacetic acid, propionic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.), and the like.

Suitable base includes an inorganic base such as alkali or alkaline earth metal hydroxide or the corresponding carbonate or bicarbonate sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, calcium 8 hydroxide, etc.), ammonium hydroxide or the like; an organic base such as an alkoxida or phenoxide of the above metal sodium ethoxide, sodium methoxide, etc.), an amine such as mono-, di or tri- alkylamine (e.g.

methylamine, ethylamine, N,N-dimethyl-l,3-pro}anediamine, trimethylamine, triethylamine, etc.) or the like.

The hydrolysis is preferably conducted under somewhat milder conditions such as under cooling or under warming in a solvent which does not have adverse influence to the reaction, e.g. water, a hydrophilic solvent such as alcohol methanol, ethanol, propanol, etc.), acetone, N,N-dimethylformamide, etc. A liquid abovementioned acid and base can also be used as a solvent.

C* Be C 0 6 Ce C C Co

CC

CC.,

eq Ce S C Ce C C .5 15 2) Reduction a•

C

e S ee S*

C

CS*

S

C

Reduction, including chemical reduction and catalytic reduction, is carried out in a conventional manner.

Suitable reducing agents to be used in chemical reduction are a metal tin, zinc, iron, etc.), or a combination of such metal and/or metallic compound (e.g.

chromium chloride, chromium acetate, etc.) and an organic or inorganic acid formic acid, acetic acid, 25 propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, etc.).

Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts (e.g.

platinum plate, spongy platinum platinum black, colloidal 30 platinum, platinum oxide, etc.), palladium catalysts (e.g.

spongy palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, etc.), or the like.

The reduction is usually carried out in a solvent such as water, an alcohol methanol, ethanol, etc.) the like.

9 The reduction is preferably carried out under somewhat milder conditions such as under cooling, at room temperature or under warming.

Process 4: Compound (I c Compound (Id) The Compound (Id) or its salt can be prepared by reducing the Compound (I c or its salt.

The reduction including chemical reduction and catalytic reduction is carried out in a conventional manner.

Suitable reducing agents may include the same as S* those exemplified in Process 3.

15 The reduction is usually carried out in a solvent such as water, an alcohol methanol, ethanol, etc.) or the like.

The reduction is preferably carried out under *somewhat milder conditions such as under cooling, at room temperature or under warming.

Process 5 SCompound (Ie) Compound Compound (I q 25 Compound (I f) and its salt can be prepared by reacting Compound (I e or its salt with Compound or its salt.

The reaction is carried out by substantially the same method of that of Process 1.

S 30 Pharmaceutically acceptable salts of the trifluoromethylketone derivatives can be prepared by a conventional method, by treating the Compound (I) with an acid or a base. Suitable examples of the acid or base may include the same as those exemplified in the explanation of "Hydrolysis" of Process 3.

10 Starting Compounds (II) to each include new compounds and can be prepared by Preparations as described hereinafter and by the similar methods thereto.

The object Compound including Compounds (I a to (I and starting Compounds (II) and (IV) include one or more isomers due to the asymetric carbon atoms and all of such isomers are included within the scope of this invention.

According to this invention, there can be obtained a mixture of diastereoisomers due to the presence of compounds bearing both R and S configurations at the chiral center marked with A of the formula as mentioned below, and there can also be obtained an optically pure compound.

It is to be noted that said optically pure compound changes to a mixture of said diastereoisomers in an aqueous and/or organic solution.

R

2

R

3 1-NHCO- )Xin-CONCHCONYCO

CF

A 3 25 .30 Further, it is to be noted that the object compound of this invention provides a hydrate form in an aqueous solution, which is included within the scope of this invention.

The trifluoromethylketone derivatives and pharmaceutical acceptable salt thereof have a human leukocyte elastase-inhibiting activity and is useful as human leukocyte elastase inhibitors for treating or preventing degenerative diseases, for example, pulmonary emphysema, atherosclerosis, rheumatoid arthritis, arthrosclerosis, osteoarthritis, psoriasis, pancreatitis, periodontosis, pulmonary fibrosis, cystic fibrosis, chronic bronchitis, bronchiectasia, diffuse 10 Starting Compounds (II) to each include new compounds and can be prepared by Preparations as described hereinafter and by the similar methods thereto.

The object Compound including Compounds (Ia) to (I and starting Compounds (II) and (IV) include one or more isomers due to the asymetric carbon atoms and all of such isomers are included within the scope of this invention.

According to this invention, there can be obtained a mixture of diastereoisomers due to the presence of cor,pounds bearing both R and S configurations at the chiral center marked with A of the formula as mentioned below, and there can also be obtained an optically pure compound.

15 It is to be noted that said optically pure compound changes to a mixture of said diastereoisomers in an aqueous and/or organic solution.

DS 6* a

O

S 4

S.

*5*6 6@ S 4 o .5 0@ S SS 0 6 a

OS

0 2 R R R R -NHCO-- \~j-'XNHHCO-Y-CONHCCCF 3 Further, j.t is to be noted that the object compound of this invention provides a hydrate form in an aqueous solution, which is included within the scope of this invention.

The trifluoromethylketone derivatives and pharmaceutical acceptable salt thereof have a human leukocyte elastase-inhibiting activity and is useful as 30 human leukocyte elastase inhibitors for treating or preventing degenerative diseases, for example, pulmonary emphysema, atherosclerosis, rheumatoid arthritis, arthrosclerosis, osteoarthritis, psoriasis, pancreatitis, periodontosis, pulmonary fibrosis, cystic fibrosis, chronic bronchitis, bronchiectasia, diffuse 11 0* g *r 0

S.

.0 0 805 0 S 5005 St 05 S *i O 0 0 *B O panbronchiolitis, respiratory injury, adult respiratory distress syndrome and the like, and further is useful for treatment or prevention of asthma, graft rejection nephritis, hydroa, disseminated intravascular coagulation, shock, systemic lupus erythematosus, clone disease, ischemia-reperfusion injury, chronic obstructive pulmonary disease (COPD), premature rupture of the membrane (PROM), corneal sarring or fibroblast proliferation (ocular coagulation, burns, mechanical and chemical injury, kerato-conjunctivitis, etc.), and sepsis.

In order to illustrate the usefulness of the trifluoromethylketone derivatives and their pharmaceutically acceptable salt, pharmacological test data thereof are shown below.

15 Test 1. Protease Inhibition assay (in vitro) Method A buffer used throughout the assay was 0.1M HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) containing 0.5M NaCl, pH 7.5. Twenty-five microliters of 2 mM methoxysuccinyl-(Ala) 2 -Pro-Val-p-nitroanilide (100 mM of dimethyl sulfoxide solution were diluted in the buffer) and 50 il of sample (10 il of sample in organic solvent was diluted 5-fold in the buffer) were mixed in wells of 96 well-microliter plate. An absorbance of the mixture in 25 wavelength at 415 nm was measured by a microplate reader (Corona Electric Co., Ibaraki, Japan). After the measurement, pl of 6 pg/ml human sputum elastase (HSE) was added and the mixture was allowed to stand for 30 minutes at room temperature. Then, the absorbance at 415 nm was measured.

30 Percent inhibition by drug was determined by 100 x inhibitor present/"r" inhibitor absent), where is absorbance after 30 minutes incubation minus absorbance before enzyme addition. Effect of inhibitors against porcine pancreas elastase (Type IV, 5 pg/ml final) was assayed similarly using N-succinyl-(Ala) -p-nitroanilide. HSE was obtained from Elastin Products Company Inc MO, U.S.A. All other substrates and protease were purchased from Sigma Chemicals Co.

d St..

.i 4 *r 0

S,

12 Inhibitory effect on several serine protease activity

IC

5 0

(M)

Test Compound Human sputum Porcine pancreas (Example No.) elastase elastase 1 4.5 x 10 7 4.4 x 10-6 -6 3 9.8 x 10 7 8.7 x 106 4 1.4 x 10 6 2.9 x 10 6 3.0 x 10-7 5.1 x 10-6 all -7 -6 6 4.3 x 10 6.9 x 10 15 7 3.2 x 10 7 3.8 x 10 6 0-7 8 8.7 x 10 1.6 x 10 9 8.7 x 10 7 9.5 x 10 1* 8.0 x 10 6 3.1 x 13 7.2 x 10 7 3.3 x 10 6 15 6.1 x 10 7 2.0 x 10 6 16 7.1 x 10-6 3.0 x 10- 17 1.1 x 10 6 3.4 x 10 6 -7 6 18 6.1 x 10 3.8 x 106 %0,046 -7 -6 19 6.8 x 10 3.0 x 10 20 8.9 x 10 7 2.4 x 10 6 -6 0-6 21 1.2 x 10 3.7 x 10 *a 22 6.8 x 10 7 4.9 x 10 6 23 8.1 x 10 7 2.4 x 10 6 t' 24 1.4 x 10- 3.8 x 10Q 6 30 27 2.4 x 10 6 1.6 x 10 29 2.4 x 10 6 1.9 x 10 13 Test 2. Determination of the activity in elastase-induced pulmonary damage.

Method Hamsters under pentobarbital anesthesia were used.

Saline or saline-containing human sputum elastase was instilled irtratracheally via a small incision in the ventral neck region using 1-ml syringe with a 27-gauge needle. After 3 hours, animals were sacrificed by CO 2 asphyxiation, each animal's trachea was reexposed. The lungs were then laveged using a 2.5-ml aliquot of saline and then withdrawing the saline, yielding a final volume of approximately 1.5 ml bronchoalveolar lavage (BAC) fluid from each animal.

15 The cells of BAL fluid were collected by centrifugation and were then diluted with distilled water to disrupt, and the hemoglobin contents determined spectrophotometrically at 541 nm.

Test drugs were dissolved in saline or methyl cell'ilose, and instilled intratracheally in the same manner as used to instill elastase, at 5 minutes before instillation of elastase.

Result Inhibitory effect on elastase-induced lung hemorrhage 5 min.

ED

5 0 Test Compound ose Hemorrhage inhipredose S(Example No.) (g/site) (OD541 nm) bition (pg/site) Normal 0.6449± 0.173*** Control 14.66 1.68 13 1 11.63 1.99 21.6 4.217 1.02** 74.5 100 0.6384± 0.222*** 100.0 14 Normal Control 19 Normal Control *15 as 255 ov 3 0 a ae Normal Control 21 Normal Control 23 1 100 1 100 1 100 10 100 10 1 100 10 1 100 0.8352± 12.89 11.59 1,663± Q. 2141± 0.8352± 12.89 10.94 2. 522 t 0. 2680± 0.6449± 14.66 12.76 5.372± 0. 6476± 0.9424± 11.05 9. 435 3 .412 0.7258± 0. 3203± 14.11 10.68 4.87 8± 0. 3451± 0. 3199± 14.11 12.05 8.155 4.545 0 .423*** 1.44 1.40 0. 690*** 0. 020*** 0.423*** 1.44 1.35 0 .803*** 0. 050*** 0 .173*** 1.68 1. 11 2.06** 0. 129*** 0. 403*** 1.40 0.941 1. 31** 0.303*** 0.159** 1.80 1.25 0.917** 0. 084*** 0.19* 1.80 1.95 1.76* 1. 49** 10.8 93.1 105.2 16.1 86.0 104.7 13.6 66.3 100.0 16.0 75.6 102.1 24.8 66.9 99.8 14.9 43.2 69.4 2.4 3.2 3.7 17.9 Normal Control 27 Normal Control 29 *P<0 .05, P<0-01, ***P<0.001 (Student's t-test) 15 Test 3: Effect on human sputum elastase induced paw edema in mice.

Materials and methods; Male C57BL mice at the age of 7-8 weeks were obtained from Japan Clear Inc,.

Human sputum elastase (HSE) was purchased from Elastin Products Company, Inc,. The test drug was administered subcutaneously, and 15 minutes later, HSE was injected into the right hind footpad at the dosage of pg/site, and saline into the left hind footpad as the control. After 2 hours of HSE injection, the paw edema was measured with the dial thickness gage, and the difference of the thickness between right and left hind 15 footpads was calculated.

@0 60

S

6

S.

6 S SWR S 0@ *e 6 So 666* ease** *6 *:eo25 0@u *0 0 9 0 06 S. r S 56 Results: Effect on the elastase induced paw edema in mice Treatment of the compound Thickness of footpad %Inhibition -2 of Example 19 n (x 1 -mm) of paw edema (mg/kg) Control 5 46.8 5.76 1 5 43.8 5.67 6.4 10 5 37.0 13.41 20.9 100 5 27.0 4.66* 42.3 p<0.05 vs control group (Student's t test) Test 4: Effect on experimentally induced emphyseman in hamsters Materials and methods: 16 Male golden Syrian hamsters, weighing approximately 120 g, were obtained from Japan SLC Inc,.

Porcine pancreatic elastase (PPE) was purchased from Elastin Products Company, Inc,. Dialferin was purchased from Japan ROche Inc,.

Hamsters were anesthetized intraperitoneally with Pentobarbital. The compound of Example 19 was dissolved in saline. Both prior art compounds A and B were suspended in 0.5 methyl cellulose. The drugs were instilled intratracheally through the oral cavi5ty, minutes before 100 pg/site of PPE in 0.2 Ml of saline instillation. Three weeks after PPE instillation, the hamsters were anesthetized with Pentobarbital.

Respiratory mechanics were studied in supine hamsters using a whole body, constant-volume, variable pressure plethysmograph to measue volume. A water-filled esophageal catheter was used to estimate pleural pressure.

Quasi-static deflation pressure-volume curves were obtained by intraperitoneally administered Dialferin to suppress spontaneous breathing inflating th e lungs to a transpulmonary pressure (PL) of 30 cm H2 0, permitting slow deflation to a PL of 0 cm H 20 and gently aspirating to a PL of -20cm H2 0. Quasi-static lung compliance (cst) was defined as the slope of the steep portion of the deflation P-V curve in the mid-volume range. Vital capacity was defined as the difference in lung volume between (volume at PL=25cm H 20) and RV-20 (volume at H Results: Pretreatment with the compound of Example 19 prevented the development of PPE-induced increases in lung mechanics in a dose dependent manner as shown in the following table. Considering Cst and VC values, the 17 potency of the compound of Example 19 was superior to the prior art compounds A Fnd B.

Effect on experimentally induced emphysema in hamsters Treatment n Cst VC (rg/site) (ml/cmH 2 O0) (ml) 0 0 00 00 a 0 0 see": 0 Exp. 1 Normal Control Compound of Example 19 1 100 Exp. 2 Normal Control Prior art Compound A 100 1000 Exp. 3 Normal control Prior art Compound B 32 320 0.53 1.54 1.39 0.70 0.53 0.51 1.62 1.55 1.22 0. 02*** 0.10 0.05(15%) 0.02(100%)*** 0.02*** 0 .12 0.11(7%) 0.15(37%) 4.9 7.3 7.0 5.8 0.1** 0.2 0.1(12%) 0.1( 0.1 0.21% 0.2(9%) 4.6 6.8± 6.9± 6. 6 0.49 1.18 1.95 0.74 0.02** 0.21 4.6± 7 .0 0.2 0.2 0.3(39%)* 0.08(32%) 0.05(63%)* 6.1 5.8 Cst=Quasi static lung compliance, VC=Vital capacity (%=Inhibition ***:pcZO.OO1 vs ContrZol (Student's t test) 18 Prior art compound A (Japanese Kokai Tokkyo Koho No.61-218518): CH CH 3

CH

I

CH

,,3 Cl- -sO, 2 NHCO- CONHCH-CON C1-S 0y

CONHCH-COCF

3 I. e ft 0 5 oo o

SO

S S

S

S.*

ee *O «I

S

'.me

S

S

o roolt 15 Prior art compound B (Japanese Kokai Tokkyo Koho No.2-256657): CH CH CH CH 3 3 CH CH Cl -SO NHCO-< \-CONHCH-CONCH 2 CONHCH- COCF 3 2 _J Pharmaceutical compositions of this invention can be used 20 in a conventional pharmaceutical forms such as powders, fine granules, granules, tablets, dragee, injections, inhalations, microcapsules, capsules, suppository, solution, suspension, emulsion, syrups and the like. If desired, diluents or disintegrators sucrose, lactose, starch, crystalline 25 cellulose, low-substituted hydroxypropyl cellulose, synthetic aluminum silicate, etc.), binding agents cellulose, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethyleneglycol, etc.), coloring agents, sweeting agents, lubricant (e.g.

magnesium stearate, etc.) or the like, may be dispensed with said composition.

19 The dosage of said composition of this invention depends on the patient's age, body weight, condition, etc., and it is generally administered by the oral or inhale route at the daily dose level of 1 mg to 1 g as the object compound or its pharmaceutically acceptable salt, preferably 10 mg to 500 mg on the same basis, at the interval of 1 to 3 times a day. Typical unit doses may be mg, 10 mg, 20 mg, 50 mg, 100 mg and the like, although these are only examples and not limitative.

SS q 000 U 0 0 00 Op The following Preparations and Examples are given for the purpose of illustrating this invention.

In the Preparations and Examples, the following abbreviations are used.

WSCD l-ethyl-3-(3-dimethylsinopropyl)carbodiimide HOBT N-hydroxybenzotriazole DMF N,N-dimethylformamide DMSO dimethylsulfoxide *000 0* 0 20 Preparation 1 To a solution of N-(tert-butoxycarbonyl)-L-valine (10.86 g) and L-proline benzyl ester hydrochloride (12.09 g) in DMF (50 ml) were added HOBT (6.76 g) and WSCD (7.76 g) under ice-bath cooling. After being stirred at room temperature for 18 hours, the reaction mixture was concentrated under reduced pressure, The residue was dissolved in ethyl acetate (400 ml) and washed with aqueous citric acid 200 ml) saturated aqueous sodium bicarbonate (200 ml). The solution was dried over magnesium sulfate and evaporated under reduced pressure to S, give N-(tert-butoxycarbonyl)-L-valyl-L-proline benzyl ester (20.06 g) as an oil.

TLC (Kiesel gel 60 F254 silica gel plate, Merck) 15 (The same meaning in the following Preparations and Examples, unless otherwise indicated) Rf 0.62 (Hexane:AcOEt 2:1) 20 0 *0 2 The following compounds were prepared by a similar method to that of Preparation 1.

Preparation 2 4-(Methoxycarbonyl)phenylcarbonyl-L-valyl-L-proline benzyl ester oil TLC Rf 0.89 (CHC1 3 :MeOH 10:1) Preparation 3 3(RS)-[[4-(Methoxycarbonyl)phenylcarbonyl]-L-valyl- L-prolyl]amino-1,1,l-trifluoro-2(RS)-hydroxy-4methylpentane mp 64-67"C TLC Rf 0.63 and 0.60 (CHC13:MeOH 10:1) Preparation 4 3(R or S)-[[4-(Methoxycarbonyl)phenylcarbonyll-Lvalyl-L-prolyllamino-l,l,l-trifluoro-2(R or S)-hydroxy-4methylpentane 21 mp 65-75 0

C

TLC 'Rf :0.65 (CHC1 3 :MeOH 1L0:1) (a]D 22: 56.230 (C=0.14, MeOI-) Preparation 3 (R or S) (Benzyloxycarbonylmethylaniinocarbonyl) phenylcarbonyl] -L-valyl-L-prolyll amino-i, 1,1-trif luoro-2 (R or S) -hydroxy-4-methylpentane mp .0TLC Rf 0.65 (CHC 3 :MeOH 10:1) [at) 22 46.59* (C=0.165, MeOH) 0..6 0 Preparation 6 3 (R or S) -[[4-(Benzyloxycarbonylme aiylaminocarb''&onyl )phenylcarbonyl 3-L-vaiyl-li-proly1 iiamino-i ,1,1trifluoro-2 (R or S) -hydroxy-4-methylpentane mp 80-9Q 0

C

TLC Rf 0.60 (CHC1 3 :MeOH 10:1) al22 :-27.591 (C=0.165, MeCH) Preparation 7 3 (R or S) £4-(1tethoxycarbonyl)phenylcarbonyJJ valyl-L-prolyllanino-i, 1, i-trifluoro-2(R or S) -hydroxy- 4-methylpentane mp :68-85'C TLC Rf 0.60 (CHC 3 :MeOH 10:1) 22 42.63* (C=0.175, MeOl) *5 0 4 50

S*

0

OS

0 *0 00 0 0 050000

S

Preparation 8 N- (tert-But~oxycarbonyl) -L-valyl-iL-proline benzyl ester (20.0 g) was dissolved in' 4N hydrogen chloride in dioxane (30 ml) under ice bath cooling. After being stirred at room temperature for one hour, the reaction mixture was evaporated9 under reduced pressure. The residue was pulveriZed with ether to give L-valyl-L- 22 proline benzyl ester hydrochloride (14.56 g).

mp 66-69°C TLC Rf 0.55 (CHC13:MeOH 10:1) Preparation 9 A solution of N-4-(methoxycarbonyl)phenylcarbonyl-Lvalyl-L-proline benzyl ester (18.53 g) in methanol (150 ml) was hydrogenated over 10% palladium on carbon (1.0 g) at 3 atmosphere pressure of hydrogen for 1.5 hours at room temperature. After the catalyst was removed by filtration, the filtrate was evaporated under reduced pressure to give 4-(methoxycarbonyl)phenylcarbonyl-Lvalyl-L-proline (14.20 g).

mp 68-71°C 15 TLC Rf 0.27 (CHC1 3 :MeOH 10:1) 00 00 *i S @0 0 000 0 *0 .0 00 00.0 00 .6 0 *0 9 *00* 0 0 Preparation To a solution of oxalyl chloride (0.82 ml) in dichloromethane (5 ml) were added dimethylsulfoxide (1.34 20 ml) and a solution of 3(RS)-[[4-(4-m5,hnoxycarbonyl)phenylcarbonyl -L-valyl-L-prolyl amino-1,l,l-trifluoro- 2(RS)-hydroxy-4-methylpentane (2.5 g) in dichloromethane (10 ml) at -70"C successively. After the mixture was stirred for one hour at -40 0 C, triethylamine (2.63 ml) was added. The mixture was stirred for an additional minutes at the same temperature and washed with hydrochloric acid (15 ml) and 5% sodium bicarbonate aqueous solution (15 ml). The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified with silica gel (50 g) column chromatography (chloroform:methanol 50:1) to give 3(RS)-[[4-(4-methoxycarbonyl)phenylcarbonyl]-L-valyl-Lprolyllamino-1,1,1-trifluoro-4-methyl-2-oxopentane (2.18 g).

mp 67-70 0

C

23 TLC Rf 0.51 (CHC1 3 MeOH 10:1) Preparation 11 To a solution of 4-'nethoxycarbonyl) phenylcarbonyll-l-valyl-L-prolyl3amino-1,1,1-trifluoro-4methyl-2-oxopentane (2.1 g) in methanol (40 ml) was added 1N aqueous sodium hydroxide (15 ml) under ice-bath cooling. After the mixture was stirred at room temperature for 2 hours, methanol was evaporated. The concentrate was washed with ether (30 ml), then acidified to p1 2 with 1N hydrochloric acid. The aqueous solution was extracted with ethyl acecate (20 ml). The extract was washed w.th brine (10 ml), dried over magnesium sulfate 0*re and concentrated under- reduced pressure to give 3 (RS) (4-carboxyphenylcarbonyl) -L-valyl-L-prolylj aminoill-trifluoro-4-methyl-2-oxopentane (1.94 mp 215-220-C TLC Rf 0.63 (CHCl 3 :eOH:AcM 8;2:1) 4.66: 20 The following compounds ware prepared by a similar method to that of Preparation 11.

*0 00 a 0 O 00e r, S 0e 0 PrCIaration 12 3(R or S)-[E4-(Carboxy)phenylarbonyl]-L-valyl-L- 25 prolyl amino-1,1,1-trifl oro-2(R or S)-hydroxy-4methylpentane (0.61 g) rr~p 266-269 0

C

TLC Rf 0.42 (Benzene:EtOAc:AcOH 20:20:1) 22 2 -49.71' (C0.l, MeOHI-) Preparation 13 3(R or S) -1 Carboxy)phenylcarbonyl]-,-valyl-Lprolyl amino\-s 1,1 txif luoro-2 (R or S) -hydroxy-4methylpentane mp 265-26 b '-24 TLC Rf 22 0.40 (Benzene:EtOAc:AcOH 20:20:1) -64.25' (C=0.16, MeOll) 05 0 0

S

0e 0 0 000 0@ S 0 *5 0e*S 0* 06 S *5 S S Preparation 14 A solution of 28% sodium methoxide in methanol ml) was added to a solution of 3(RS)-amino-1,1,1tritluoro-2 (RS) -hydroxy-4-methylpentane hydrochloride (3.2 g) in ethanol (30 ml) at room temperature. After removal of the precipitat".ed sodium chloride by filtration, (2R,3R)-L-tartaric acid (2.3 g) was added to the filtrate.

The mixture was warmed until tartaric acid was dissolved and filtrated. The filtrate was allowed to stand for hours at room temperature. The precipitated crystalline solid (1.94 g) was collected by filtration and dissolved 15 in 1N aqueous sodium hydroxide. The solution was extracted with ethyl acetate (10 ml) and the extract was mixed with 4N-hydrogen chloride in ethyl acetate. After removal of ethyl acetate, the residue was pulverized with diisopropyl ether (10 ml) to give 3(R or S)-aniino-1,1,1- 20 trifluoro-2(R or S)-4-methylpentane hydrochloride (1.03 it00 S S SS S S. S

S

S 55 0

S*.S@

mp 165-170 0

C

TLC Rf :0.50 (CHCl 3 :MeOH 10:1) lo]22 +11.39* (C=0.13, MeOH) SeeS S S a.

S

a S 00 0 0

S

The following compound was prepared by a similar method to that of Preparation 14.

Preparation 3(R or S)-Amino-1,1,l-trifluoro-2(R or S)-4methylpentane hydrochloride mp 165-170*C TLC Rf 0.55 (CHC 3 :MeOH =10:1) a322 -10.560 (C=1.05, MeOH) 25 Preparation 16 To a solution of methyl p-aminobenzoate (0.4 g) in tetrahydrofuran (10 ml) was added trichloromethylchloroformate (0.31 g) and the mixture was allowed to stand overnight at room temperature, L-Valine benzyl ester (0.54 g) was added to the solution and the pH of the mixture was neutralized with triethylamine. After being stirred for 30 minutes at room temperature, the mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (10 ml). The extract was washed with lN-hydrochloric acid (10 ml) and aqueous sodium bicarbonate (10 ml) and concentrated to dryness to give 4- methoxycarbonyl)phenylaminocarbonyl-Lvaline benzyl ester (1.17 g).

15 TLC Rf 0.55 (CHC1 3 :MeOH 10:1, V/V) a.

0o a 20 Preparation 17 4-(Methoxycarbonyl)phenylaminocarbonyl-L-valine (0.83 g) was prepared from 4-(methoxycarbonyl)phenylaminocarbonyl-L-valine benzyl ester (I g) by a similar method to that of Preparation 9.

TLC Rf 0.3 (CHC1 3 :MeOH:H 2 0 65:25:4) Oil Preparation 18 [4-(Methoxycarbonyi)phenylaminocarbonyll-L-valyl-Lproline benzyl ester (1.0 g) was prepared from 4-(methoxycarbonyl)phenylaminocarbonyl-L-valine (0.83 g) and L-proline benzyl ester hydrochloride (0.65 g) by a similar method to that of Preparation 1.

TLC Rf 0.60 (CHC1 3 :MeOH 10:1) Oil Preparation 19 (4-(Methoxycarbonyl)phenylaminocarbonyl]-L-valyl- 26 L-proline (0.69 g) was prepared from [4-(methoxycarbonyl)phenylantinocarbony1] -L-valyl-L-proline benzyl ester g) by a similar method to that of Preparation TLC Rf 0.35 (H1 MeOH:H 2 0 65:25:4) Oil Preparation 3 (RS) (Methoxycarbonyl) p)aenylaminocarbonyl IL valyl-L-propyllamino-l,l,l-trifl'aoro-2(RS) -hydroxy-4methylpentane (1.03 g) was prepared from @6 (methoxycarbonyl lphenylaiinocarbonyl] -L-valyl-L-proline (0.69 g) and 3(RS)-amino-l,l,l-trifluoro-2(RS)-hydroxy-4- 660 methylpentane hydrochloride (0.37 g) by a similar method to that of Preparation 1.

TLC Rf 0.45 (CHC 3 :MeOH 10:1) a 0oil 20 a 6O*4 9 @0 6 S. 0

SS

*6 9 00 ii 9 6 *0w*O* S Preparation 21 3 (RS) (Methoxycarbonyl) phenylaminocarbonyl3 -Lvalyl-L-prolyl) amino-l, i, l-trifluoro-2-oxo-4-methylpentane (0.98 g) was prepared from 3(RS)-[[4-(methoxycarbonyl)phenylaminocarbonylll-L-valyl-L-prolyllamino-1,1, 1trifluro-2 (RS) -hydroxy-4-methylpentane (1.0 g) by a similar method to that of Preparation nip 90-100 0

C

TLC Rf 0.50 (CHC 3 :MeOH 10:1) 0 6 Preparation 22 3 (Rs) -t[4-(Carboxy)phenylaminocarbonyl]-L-valyl-Lpro-.,yl~ainino-1,1,1-trifluoro-2-oxo-4-methylpentane (0.2 g) ws prepared from 3 (RS) (methoxycarbonyl)phenylaminocarboniyl) -L-valyl-L-prolyllamino] l-trifluoro-2-oxo-4methylpentane (0.3 g) by a similar method to that of Preparation 11.

nip 125-1301C 27 TLC Rf 0. 50 (CHCl 3 :MeQH: H 2 0 65:25:4) 0e 9 4

S.

@0 0 0 S0 S S

S.

OSS*

9 OS S

OS

0O S *0 Preparation 23 To a solution of N-(tert-butoxycarbonyl)--L-valine (4.35 g) and triethylamine (2.13 g) in dry CH 2 Cl 2 (40 ml) was added isobutyl chioroforiate (2.87 g) at -20 0 C. After being stirred at same temperature for 30 minutes, a solution of N-(2-indanyl)glycine benzyl ester (5.37 g) in dry CH 2 C1 2 (20 ml) was added at -20 0 C. The reaction mixture was stirred at -10 0 C for one hour then at room temperature for 4 hours. After the reaction mixture was evaporated under reduced pressure, the residue was dissolved in ethyl acetate (100 ml) and washed with saturated aqueous sodium bicarbonate (100 ml x The 15 organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified with silica gel (50 g) column chromatography (CHC 3 :AcO~t =10:1) to give N-(tert-butoxycarbonyl)-Lvalyl-N-(2-indanyl)glycine benzyl ester (2.20 g) as a~n 20 oil.

40040: a 0050 9 0* *5 0 S. S 5* *5 0

S

0O S. 0 a i.9950 9 0 TLC Rf 0. 78 (Hexane: AcOEt 2: 1) Preparation 24 L-Valyl-N- (2-indanyl) glycine benzyj ester hydrochloride (1.83 g) was prepared from N- (ter t-butoxyc arbonyl) -I,-valyl--N- 2-indanyl) glycine benzyl ester (2.14 g) by a similar method to that of Preparation 8.

mp 162-163 0

C

TLJC RE 0.58 (CHCl 3 :MeOH =10:1) Preparation N-[t4- (Methoxycarbonyl) phenylcarbonyl]3-L-valyl-'N- (2indanyl)glycine benzyl. ester (1.24 g) was prepared from L-valyl-N-(2-indanyl)glycine benzyl ester hydrochloride 28 (1.80 g) and terephthalic acid mono methyl ester (0.86 g) by a similar method to that of Preparation 1.

mp 72-76'C TLC Rf 0.29 (CHCl 3 Preparation 26 N- (Methoxycarbonyl) phenylcarbonyl I-L-valyl-N-(2indanyl)glycine (0.83 g) was prepared from N-C 4- (methoxycarbonyl)phenylcarbonylI-L-valyl-N- (2indanyl)glycine benzyl ester (1.20 g) by a similar method to that of Preparation 9.

mp :163-164*C TLC Rf :0.48 (CHC1 3 :MeOH 10:1)

U.

6O 0 S S *e 0 GUS 0 0 0 0.

*peS 15 Ge B

C.

0 0* 20 a 005C *0 *5 S S S Ce 0 a Preparation 27 3 (RS) I 4-(Methoxycarbonyl)phenylcarbonyl] -L-valyl- 2-indanyl~glycyllamino-1,1,1-trifluoro-2(RS)-hydroxy-4rethylpentane (1.06 g) was prepared from N-[C4- (methoxycarbonyl)phenylcarbonyl) -L-valyl-N- (2indanyl)glycine (0.80 g) and 3(RS)-amino-l,1,1-trifluoro- 2(RS"-biydroxy-4-methylpentane hydrochloride (385 mg) by a similar method to that of Preparation 1.

mp 76-78 0

C

TLC Rf 0.71 (CHC 3 :MeOH =10:1) Preparation 28 3 (RS) C 4- (IMethoxycarbonyl) phenylcar~bonylI -L-valyl-N- (2-indanyl)glycyl3amino-l,l,l-trifluoro-4-methyl-2oxopentane (0.84 g) was prepared from 3(RS)-E[4-(methoxycarbonyl)phenylcarbonyl) -L-valyl-N- (2-indanyl) glycyl] amino-l,l,l-trifluoro-2(RS) -hydroxy-4-methylpentane (1.03 g) by a similar me thod to that of Preparation mp 62-64'C TLC Rf 0.74 (CHCl 3 :MeOH -1 10:1) 29 Preparation 29 3(RS)-[(4-Carboxyphenylcarbonyl)-L-valyl-N-(2indanyl)glycyl3amino-1,1,1-trifluoro-4-methyl-2-oxopentane (0.76 g) was prepared from 3(RS)-[[4-(methoxycarbonyl)phenylcarbonyll-L-valyl-N-(2-indanyl)glycyllamino-l,1,1trifluoro-4-methyl-2-oxopentane (0.83 g) by a similar method to that of Preparation 11.

mp 84-86'C TLC Rf 0.15 (CHCl 3 :MeOH 10:1) Example 1 To a mixture of glycine benzyl ester p-toluenesulfonate (66 mg) and 3(RS)-[(4-carboxyphenylcarbonyl)-Loeeo *valyl-L-prolyl amino-l ,1,1-trifluoro-4-methyl-2-oxopentane (100 mg) in DMF (6 ml) were added HOBT (26 mg) and WSCD mg) under ice-bath cooli-g. After being stirred at room temperature for 4 hours, the mixture was concentrated under reduced presrure. The residue was dissolved in ethyl acetate (30 ml) and washed with 5% aqueous citric 20 acid (20 ml), water (20 ml), 5% aqueous sodium bicarbonate ml) and brine (20 ml). The solution was dried over magnesium sulfate and evaporated under reduced pressure to give 3 (RS) t 4- E (benzyloxycarbonyl)methylaminocarbonyl) phenylcarbonyl3-L-valyl-L-prolyl amino-l ,1,1-trifluoro-4methyl-2-oxopentane (125 mg).

mp 77-78*C TLC Rf 0.56 (CHCl 3 :MeOH 10:1) The following compounds were prepared by a similar method to that of Example 1.

Example 2 3(RS)- [4-[[2-(4-Morpholinolethyllaminocarbonyllphenylcarbonyl) -L-valyl-L-prolyl] amino- 1,-trifluoro-4methyl-2-oxopentane 30 mp 98-102*C TLC Rf 0.24 (CHCI 3 :MeOH 10:1) Example 3 3 (RS) (3-Benzoylmethoxycarbonyl)propylaminocarbonyllphenylcarbonylj -L-v~ilyl-L-prolyl) amino-i, 1,1trifluoro- 4-methyl-2-oxopentcane mp 70-73*C TLC Rf 0.71 (CHC1 3 :MeO! *8 je a

S

S.

a S a

S.

a a *5 S 5 15 q. S 6O

S

*5 20 a .055 *0

S

Example 4 (4-Mdorphoino)aminocarbonyl]phenylcarbonyll-L-valyl-L-proyl amino-i, 1, 1-trif.uoro-4methyl-2--oxopentane mp 191-193'C TLC Rf 0.59 (CHC1 3 :MeQH 10:1) Example~ 3 (RS) (4-Chlorobenzyl)aminoca,,bonyl3phenylcarbonyl)-L-valyl-L-prolyllamino-1,1, 1-trifluoro-4methyl- 2-oxopentane mp 87-89'C TLC Rf 0.43 (CHCl 3 :MeQH 10:1) SB S a aS 04 S 0 25 Example 6 3 (RS) (4-Nitrobenzyl) aminocarbonyll phenylcarbonlt13 -L-valyl-L-prolyl) amino-i, 1, 1-trifluoro-4methyl- 2-oxopentane mp 94-96*C TLC RE 0.57 (CHC1 3 :MeOH 10:1) 'Example- 7 3 (Rs) (4-Chlorophenyl) aminocarbonyl3 phenylcarbonyll -L-valyl-L-prolyll amino-1, 1, 1-trifluoro-4methyl- 2-oxopentane 31 mtp t 71-72*C TL~C Rf 0.79 (CHCl 3 :MeOH 10:1) Example 8 3(RS)-t[4-[(1I(R)-Benzyloxycarbonyl)-2-pheyl)ethylaminocarbonyl 3phenylcarbonyl 3-L-valyl-L-prolyl I)amino- 1,1, 1-trifluro-4-methyl-2-oxopentane mp :65-67'C TLC Rf t 0.82 (CHC 3 :MeOH 10:1) :Its9 0 19 o *4 4 O Example 9 3(RS')-E t4-1[ (l(S)-Benzyloxycarbonyl)-2-phenylethylaminocarbonyllp~ienrvylcarbonyl3 -L-valyl-L-prolyl Iamino- MP 108-11,0 0

C

TLC Rf 0.82 (CH1 C 3 :IMeOH =10:1) ExampLe 3(RS)-t[4-tE1(S),3-bistBenzyloxycarbonyl)propyl]aminocarbonyljphenylcarbony. 3-L-valyl-L-prolyl3 amino- 1,1, 1-trifluoro-4-mnV-thyl-2-oxopentane mp :58-60 0

C

TLC RE 0.83 (CHCl 3 :MeOH 10:1) 0*04 4 04 is 400 Example 11 To a solution of oxalyl chloride (0.09 ml) in dichioromethane (2 ml) were added dimethylsulf oxide (0.15 ml) and a solution of 3(R or (benzyloxycarbonylmethylaminocarbonyl )phenylcarbonyl 3-Lvalyl-L-prolyllamino-1,1,1-trifluoro-2 (R or S) -hydroxy- 4-methylpentane (0.35 g) in dichioromethane (4 ml) at successively. After the mixture was stirred for one hour at -40 0 C, triethylamine (0.29 ml) was added. The mixture was stirred for an additional 30 minutes at the same temperature and washed with 0.5N hydrochloric acid (15 ml) 32 and 5% sodium bicarbonate aqueous solution (15 ml). The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified with silica gel (20 g) column chromatography (Chlorofozm:methanol 50:1) to give 3(R or (benzyloxycarbonylmethylaminocarbonyl)phenylcarbonyll-Lval-l-L-prolyllamino-1,1-trifluoro-2-oxo-4methylpentane (0.22 g).

mp 159-161 0

C

TLC Rf 0.63 (CHC.1 :MeOH 10:1) 223 V* raD -39.66" (C=0,1.05, MeOH)

D

a a@ The following compound was prepared by a similar Ba method to that of Example 11.

~WG4 *e Example 12 3(R or $)-[[4-(Benzyloxycarbonylmethylaminocarbonyl)phenyloarbonyll-L-valyl-L-prolyllamino-1,1,1-trifluoro-2oxo-4-methylpentane n 20 mp 141-143 0

C

a:TLC Rf 0.68 (CHC1 :MeO) r 10:1) it 0 22 3 [laD: -41.90- (C=0.15, MOH) a a Example 13 a a A solution of 3(RS)-114-E(benzyloxycarbonyl)methylaminocarbonyl phenylcarbonyl l-L-valyl-L-prolyl amino-1,1,1-trifluoro-4-methyl-2-oxopentane (70 mg) in a a mixture of methanol (10 ml) and water (1 ml) was hydrogenated over 10% palladium on carbon (1.0 g) at 3 atmosphere pressure of hydrogen for 3 hours at room temperature. After the catalyst was removed by filtration, the filtrate was evaporated under reduced pressure to give 3(RS)-[4-(carboxymethylaminocarbonyl)phenylcarbonyll-L-valyl-L-prolyllamino-1,1,1-trifluoro-4methyl-2-oxopentane (61 mg).

33 mp 99-103'C TLC Rf 0.17 (CHCl 3 :MeOH:AcOH 8:1:1) Example A4 To a solution of 3(RS)-C[4-[(3-benzoylmethoxycarbonyl)propylaminocarbonyl1phenylcarbonyl)-L-valyl-Lprolyllamino-1,1,1-trifluoro-4-methyl-2-oxopentane (120 mg) in acetic acia (5 ml) was added'zinc powder (120 mg) under ice-bath cooling. After the mixture was stirred at room temperature for 2 hours, zinc was removed by *0 filtration. The filtrate was evaporated under reduced pressure to give 3(RS)-[[4-C(3-carboxypropyl)aminocarbonyll phenylcarbonyl) -L-valyl-L-prolyl] amino- 1,1,1-trifluoro-4-methyl-2-oxopentane (95 mg).

mp 84-86 0

C

TLC 'Rf 0.32 (CHC 3 :MeOH:AcOH 8:1:1) 04 a 0 The following compounds were prepared by a similar method to that of Example 13.

Example 3(R or s)-[[4-(Carboxymethylaminocarbonyl)phenylcarbonyll -L-valyl-L-prolyl) amino-i, 1, 1-trifluoro-2oxo-4-metbylpentane mp 75-120 0

C

TLC (RP-18 WF 254 S (made by E. Merck) Rf 0.55 (MeO:H 2 0 22 -35.100 (C=0.105, MeGH) Example 16 3(R or S)-C[4-(Carboxymethylaminocarbonyl)phenylcarbonyll-L-valyl-L-prolyllamino-i,1,1-trifluoro-2oxo-4-methylpentane mp 90-1100C TLC (RB-18 WF 25 4 S (made by E. Merck) 34 Rf 0.50 (MeOH:H 2 O 22 CalD 50.Q4o (C=0.115, MeOH) Example 17 3(RS)-[[4-[(l(S),3-Dicarboxypropyl)aminocarbonyllphenylcarbony.3-L-valyl-L-prolyliamino-i,1,1-trifluoro- 4-methyl-2-oxopentane mp 74-76CC TLC Rf 0.13 (CHCl 3 :MeOH:AcOH 8:1:1) s 55

S

S

5* 5

S

OS

S

00

OS

OS S S .0 Example 18 A solution of 3(RS)-[[4-E(4-nitrobenzyl)aminocarbonyl I phenylcarbonyl I -L-valyl-L-prolyl 3 amino- 1,1, 1-trifluoro-4-mnethyl-2-oxopentane (50 mg) in methanol (10 ml) was hydrogenated over 10% palladium on carbon mg) at 4 atmosphere pressure of hydrogen for 2 hours. The catalyst was removed by filtration and the filtrate was evaporated under reduced pressure to give aminobenzyl) aminocarbonyl I phanylcarbonyl 3 -L-valyl-t- 20 prolyllamino-l1l,1-trifluoro-4-methyl-2-oxopentane (46 mg).

S

0 5* 5 00 mp 90-92*C TLC Rf 0.44 (CHC13:MeOH:AcOH 8:1:1) Example 19 To a solution of 3(RS)-[[4-(carboxymethylaninocarbonyl) phenylcarbonyl-L-valyl-L-prolyl)amino-i,1,1trifluoro-4-methyl-2-oxopentane (0.50 g) in water (10 ml) was added 1N aqueous sodium hydroxide (0.88 ml) at room temperature. The solution was lyophilized to give sodium salt of starting material (0.52 g).

mp >230 0

C

TLC Rf 0.17 (CHCl 3 :MeOH:AcOH 8:1:1) The following compound was prepared by a similar 00 S S q S 90 Os 4~ 000 0

~S

0 0 50 0500 50 OS S 00 OS 0 00 0 0~S ~0 0 0~ 55 55 0 S. 0 0i

S

OSSOSe

S

@000 o 05 00 0 050000 0 method to that of Example 19.

Exampl..e A sodium salt of 3(RS)-[[4-f(3-c!-rboxypropyl)aminocarbonyllphenylcarbonyll -L-valy~l-L-proiyl Iamino- 1,1 ,1-trifluoro-4-methyl-2-oxopentane mp 06-69*C TLC Rf 0,32 (CHCl 3 :MeOHzACOH =8:1:1) 10 Example 21 16a solutio~n 0. 3(RS)-[E4-1t2- 4-morpholino)ethyamitocarbonyl jheny~Lcarbony1) -L-valyl-L-prolyl) amino- 1,1,1-trifluoro-4-iMethyl-2-oxopentane (80 mg) in 1,4-dioxane (1 ml) was added 4N-hydrogen chloride in aioxane (0.1 ml). The mixture was stirred at room temperature for 10 minutes, and evaporated to give 3(RS)-Et,4-[t2-(4-morphiolino)ethylaninocarbonyl3phenylcarhonyl) -L-valy1-L-prolyli amino-ie,l,-trifluoro-4methyl-2-oxopentane hydx~ochloride (83 mg).

mp 4. 64-650C T2LC Rf :0.24 (CHC1 3 ;MeOH =10:1) Example 22 3 (RS) C[4- (Benzyloxycarbonylnidtlylaminocarbonyl) 25 phenylaminocarbonyll -L-valyl-L-prolyljamino-l,L 1trifluoro-2-oxo-4-methylpefttane (0.25 g) was prepared from 3 (RS) 4- (carboxy)phernylaminocarbonyl3 -L-valyl-L-ptolyl) amino-)., ),-trifluoro-2-oxo-4-methylpentane (0.2 g) and glycine benzy. ester para-to2.uenesulfonate (0.13 g) by a similar method to that of Example 1.

nip 65-70'C TLC Rf :0.15 (CHC 3 :MeOH =10:1) Example 23 3 (RS) (Carboxymethylaminocarbonyl iphenylamino- 36 .0 0 00 0 0 *0 06 so0 carbonyl I-L-valyl-L-proJyl Iamino- 1,1, 1-trif luoro- 2-oxo- 4methyl.pentane (0.1L4 g) was prepared from 3 (RS) I[4- (benzyloxycarbonylmethylamiiocarbonyl) phenylaminocarbonyl) I-L-valyl-L-prolyi] amino-i, trifluoro-2-oxo-4-inethylpentane (0.2 g) by a similar method to that of Example 13.

nip 1 98-1280C TLC Rf :0.25 (CHC1 3 :MeOHflU 2 0 65:25:4) 10 Exam*Ple 24 3 (RS (Dimethylaminocarbonylmethylaminocarbonyl) phenylaniinocarbonyl] -L-valyl-L-proly. Iamino-i, 1,1trifJluoiro-2-oxo-4-methylpentane (0.03 g) was prepared from 3 (Rs) -EJ4- (carboxymethylaminocarbonyl) phenylaminocarbonyil)-L-valyl-L.-prolyllamino-1, 1, J-trifluoro-2-oxo-4methylpentane (0.08 g) and dimethylamine hydrochloride (0,012 g) by a similar method to that of Example 1.

m7? 115-125 0

C

T3LC Rf :0.20 (CHC 3 :MeOH =10:1) 0 900000 0 *o00 0 00 00 0 @9 0 00 00 0 @90900 9 9 SO 00 @0 @0 0 .00.0: 25 0 Example 3 (RS) -([4-[£(Benzyloxycarbonyl )methylaminocarbonyl) phenylqarbonylI -t-va3.yl-N- (2-indanyl) glycyl) amino-1,1,ltrifluoro-4-methyl-2-oxopentane (250 mg) was prepared from glycine benzyl ester p-toluenestulfonate (118 mg) and 3 (RS) (4-carboxyphenylcarbonyl)-L-vay-N-( 2-indanyl) glycyl]amino-l,.ll-trifluoro-4-methy,-2-oxopentane (206 mg) by a similar method to that of Example 1.

mp 64-66'C TLC Rf 0.81 (CHC 3 :MeOH 10:1) White powder Example 26 3 (Carboxymnethylaminocarbonyl) phenylcarbonyl) -L-valyl-N-( 2-indanyl)glycyllamino-1,3.,1- -37 triiuluoro-4-methyl-2-oxopentane (195 mg) was prepared from 3 (RS) -t E4-E (benzyl.oxycarbonyl)methylamiAnocarbonylpl~enyl~carbpnyl I -L-vjy'-N_ 2-indanyl) glycyl]I amino- 1,1,l-,trifluoro-4-methyl-2-oxopentane (22,0 mg) by a similar method to that of Example 13.

nip 88-91 0

C

TLCR 0.25 (CHC1 :MeOH:AcOH 16:1:1) aa Ga a a 10 a a a a a a.

a a aa~a a a 0. 0 a.

a a a.

15 a a.

a a a Ga S a. a a a a a.

a

S

a a a a a a. a a 25 a a. a a Eample 2?1 A spdiur, salt of I (cabxymethylaiinocarboniyl,)phenylcarbonyl) -L-valyl-1,-- (2-indanyl) glycy4) lamino-l,1,1-trifluoro-4-methyl-2-'oxopentane (160 wi wa~s prepaxed by a similar method to that of~ Fxample 19.~ inp :203-205 0

C

TLC Rf 0*25 (HCL 3 zYieOH:2\cOH =16:1:1) E xample 28 3(RS)-[j4-t2-(4-Morpholino)e 4 -,yjaminacarbonyllphenyloarbonyl) -L-valyl-N- (2 indanyl) glycyl] amino-i 1,1trifl1uoto'-4-methyl-2-oxopentane (180 mig) was prepared from 4 2 -aminopth1) morpho ine (46 mg) and 3 MRS) t(4,-carboxyphenylcarbonyl) 2-inidanyl) glyoyllami,,io-,,-trilu:oro-4-methyl-2-oxopentane (206 mrg) by a timilar r tetbod to that of Example- 1.

mp :76-80o(0 TLC RE 0. 38 (C1{C1 3 :MeO1= 10:1) Example 29 A hydrochloride of 3(RS)-Et4-EE2-(4-morpholino)ethyl) aiinocarbonyllphenylc~ rbonyl) -L-valyl-N- l-indanyl) glycyl~amino-l1,l-trifluoro-4-methyl-2-oxopent'ane (17b mg) was prepared by a similar method to that c'f Example 21.

nip :96-99,C, TLC Rf 0.38 (CEC1 3 :MeOH 10:1) 38 Example 3(RS)- £4-(ethoxycarbonyl)methylaminocarbonyl phenylcarbonyll-L-valyl-L-prolyl]amino-1,1,1-trifluoro-4-methyl- -2-oxopentane was prepared by a similar method to that of Example 1.

mp: 96-99 0

C

10 a 60 0 5 9* 0 60 *b 0.; *0g 25

S

20 *0 0 25 00j Example 31 To a solution of 4-(ethoxycarbonyl)methylaminocarbonyl 3 phenylcarbonyl3 -L-valyl-L-'prolyl 3 amino- 1,1,-tAifluoro-4-methyl-2-oxopentane (5.0 g) in methylene chloride (60 ml), methanol (10 ml) and water ml) was added aqueous sodium hydroxide (NaGH 0.6 g in water (5 ml)) under ice-bath cooling. After the reaction xixture was stirred at 0-10w0 for 10 minutes, it was adjusted to pH 9 with 6N hydrochloric acid and then aqueous solution was washed with methylene chloride ml). To the aqueous solution were added sodium chloride (5 g) and ethylacetate (60 ml), and then the mixture was acidified to pH 2 with 6N hydrochloric acid. Ethyl acetate solution was washed with brine (30 ml), dried over magnesium sulfate and concentrated under reduced pressure to the volume oi 20 ml. Thei concentrated solution was added dropwise to isopropyl ether (225 ml) at room temperature. Precipitate was filtered and then dried to give 3(RS)-lt4-(carboxymethylaminocarbonyl)phenylcarbonyll-L-valyl-.L-prolyl3amino-i,1,1-trifluoro-4-iethyl- 2-oxopentane (3.24 g).

mp: 99-1030C

Claims (16)

1. New trifluoromethylketone derivatives of the following formula R 2 R 3 01 1 R 1-HCO- [n-CONHCHCO-Y-CONHCHCOCF 3 wherein R is lower alkyl which has one or two substituents selected from carboxy, esterified carboxy and di-lower alkylcarbamoyl; phenyl(lower)aikyl, the phenyl moiety of which may have halogen or nitro or amino and the altyl moiety of which may have carbox2 or esterified carboxy; halo-phenyl; morpholino; or morpholiino(lower)alkyl, R 2 and R 3 are each lower alkyl, X is -NH-, n is 0 or 1, and Y is -N or -NCH 2 and pharmaceutically acceptable salt thereof.

2. New trifluoromethylketone derivatives according to 6claim 1 which are represented by the following formula 30 R R -NHC01 R R"CO- -OCONHCHCON ~CONHCHCOCF 3 40 10 S OS 15 wherein R is lower alkyl which has one or two substituents selected from carboxy, esterified carboxy and di-lower alkylcarbamoyl; phenyl(lower)alkyl, the phenyl moiety of which may have halogen or nitro or amino and the alkyl moiety of which may have carboxy or esterified carboxy; halo-phenyl; morpholino; or maorpholino(lower)alkyl, and R 2 and R 3 are each lower alkyl, and pharmaceutically acceptable salt thereof.

3. New trifluoromethylketone derivatives according to claim 2, wherein 1 R is lower alkyl which has carboxy or esterified carboxy, and 2 3 R 2 and R are each lower alkyl.

4. New trifluoromethylketone derivatives according to claim 3, wherein R is carboxymethyl, and R 2 and R 3 are each isopropyl. New trifluoromethylketone derivatives according to claim 4 which are 3(RS)-[[4-(carboxymethylamino- carbonyl)phenylcarbonyl -L-valyl-L-prolyl amino- 1,1,l-trifluoro-4-methyl-2-oxopentane.

6. New trifluoromethylketone derivatives according to claim 4 which are a sodium salt of carboxymethylaminocarbonyl)phenylcarbonyl]-L- valyl-L-prolyllamino-,1,l-trifluoro-4-methyl-2- oxopentane.

7. A process for preparation of trifluoromethylketone derivatives of the following formula or their salt 0 20 S 20 **205 S 25 41 R3 wherein R1 is lower alkyl. which has one or two substituents selected from carboxy, esterified carboxy and di-.lower alkylcarbamoyl; phenyl (lower) alkyl, the phenyl moiety of which may have halogen or nitro or amino and the alkyl moiety of which may have carboxy or esterified carboxy; hala-phenyl; morpholino; or morpholino (lower) alkyl, R 2 and R 3 are each lower alkyl, X is -NH-, n is 0 or 1, and Y is or l -NCH 2 which comprises reacting a compound of the following formula or its salt 3 3 with a compound of the formula R 1 -NH 2 or its salt, or oxidizing a compound of the following formula or its salt: e. a a. a. S i a. a S a a a. a a *e S. *5 a a. a a. a. a. a. a a a a a a a a. a a.. a a a. S a a aesa X QHHCO-Y/\-(l -CON HCHCHCFuriki~tur 3 Uri ,or subjecting a compound of the following formula or its salt to de-;esterification reaction: 42 R 2 R 3 C I CO<I I Ra-NHCO- n- CONHCHCO-Y-CONHCHCOCF 3 reducing a compound of the following formula or its salt R 2 NO- -CHNHCO- -CONHCHCON7 R 3 CONHCHCOCF 3 or reacting a compound of the following formula or its salt 7 R3 R' R HOOC-R -NHCO- CONHCHCO-Y-CONHCHCOCF 3 or with a compound of the formula (Rd) 2 NH or its salt. In the above formulae, R is mono- or di- esterified 1 1a carboxy(lower)alkyl, R is lower alkylene, Rd is lower alkyl, and R 1 to R 3 X, Y and n are each as defined above.

8. A process according to claim 7, wherein 1 R is lower alkyl which has carboxy or esterified- carboxy, 2 3 R and R are each lower alkyl, n is 0 and Y is -N'

9. A process according to claim 7, wherein R is carboxymethyl, R 2 and R 3 are each isopropyl, n is and Y is -N 25 30 o *g* 0 0 0 0000" 0* 43 *4 49 too a 4* 0 .6 0 A process according to claim 7, wherein the object compound is 3(RS)-[[4-(carboxymethylaminocarbonyl)- phenylcarbonyl]-L-valyl-L-prolyl]amino-1,1,1- trifluoro-4-methyl-2-oxopentane.

11. A process according to claim 7, wherein the object compound is a sodium salt of carboxymethylaminocarbonyl)phenylcarbonyll-L-valyl- L-prolyl]amino-1,1,l-trifluoro-4-methyl-2-oxopentane.

12. A pharmaceutical composition comprising as an effective ingredient new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt in association with a pharmaceutically acceptable carrier or excipient.

13. A human leukocyte elastase inhibitor comprising as an effective ingredient new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt in association with a pharmaceutically acceptable carrier or excipient.

14. A pharmaceutical composition for treatment or prevention of degenerative diseases such as pulmonary emphysema, atherosclerosis, rheumatoid arthritis, arthrosclerosis, osteoarthritis, psoriasis, pancreatitis, periodontosis, pulmonary fibrosis, cystic fibrosis, chronic bronchitis, bronchiectasia, diffuse panbronchiolitis, respiratory injury, and adult respiratory distress syndrome comprising as an effective ingredient new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt in association with a pharmaceutically acceptable carrier or excipient. 4 *44443 44 ok o 20 ago i- C C ai 9 3 99 15 a *4 Sc J S 'esee. A pharmaceutical composition for treatment or prevention of graft rejection comprising as an effective ingredient new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt in association with a pharmaceutically acceptable carrier or excipient. 16, A pharmaceutical composition for treatment or prevention of nephritis, sepsis, hydroa, disseminated intravascular coagulation, shock, systemic lupus erythematosus, clone disease, ischemia-reperfusion injury, chronic obstructive pulmonary disease (COPD), premature rupture of the membrane (PROM), corneal sarring or fibroblast proliferation (ocular coagulation, burns, mechanical and chemical injury, kerato-conjunctivitis, etc.), or asthma comprising as an effective ingredient new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt in association with a pharmaceutically acceptable carrier or excipient.

17. A method of treating or preventing degenerative diseases such as pulmonary emphysema, atherosclerosis, rheumatoid arthritis, arthrosclerosis, osteoarthritis, psoriasis, pancreatitis, periodontosis, pulmonary fibrosis, cystic fibrosis, chronic bronchitis, bronchiectasia, diffuse panbronchiolitis, respiratory injury, and adult respiratory distress syndrome in a subject in need thereof which comprises.administering to the subject an effective amount of new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt. 45

18. A method of treating or preventing graft rejection in a subject in need thereof which comprises administering to the subject an effective amount of new trifluoromethylketone derivatives as defined in claims 1 to 6 or their pharmaceutically acceptable salt.

19. A method of treating or preventing nephritis, sepsis, q, hydroa, disseminated intravascular coagulation, shock, systemic lupus erythematosus, clone disease, ischemia-reperfusion injury, chronic obstructive Vpulmonary disease (COPD), premature rupture of the membrane (PROM), corneal sarring or fibroblast proliferation (ocular coagulation, burns, mechanical and chemical injury, kerato-conjunctivitis, etc.), or asthma in a subject in need thereof which comprises administering to the subject an effective amount of new trifluoromethylketone derivatives as defined in 4 0, claims 1 to 6 or their pharmaceutically acceptable 20 salt. S. Use of a compound of any of claims 1 to 6 and their pharmaceutically acceptable salts for the manufacture of a medicament for therapeutic treatment or prevention of degenerative diseases such as pulmonary emphysema, chronic bronchitis, atherosclerosis, rheumatoid arthritis, osteoarthritis, psoriasis, pancreatitis and adult respiratory distress syndrome, and nephritis, sepsis, asthma or graft rejection. 46

21. Compounds of formula processes for their preparation or pharmaceutical compositions or methcds of treatment involving them, substantially as hereinbefore described with reference to the Examples. DATED this 26th day of July, 1993. FUJISAWA PHARMACEUTICAL LTD. By its Patent Attorneys DAVIES COLLISON CAVE a a a. a a a a a a. a a a. a a. a a a. a. a. a a a. a 4~ a. a. a. a a a a a a. a a a a. a. a a a a.~ a a a. a. a 0* a A a ~O ,II 93G721,p:\oper~dab,898S3-91.2046 ABSTRACT New trifluoromethylketone derivatives of the following formula R 2 R 3 R -NHCO-_ X-CONHCHCO-Y-.CONHCHCOCFut 3 where in Ris lower alkyl which has one or two substituents selected from carboxy, esterified carboxy and di-lower alkylcarbamoyl; phenyl~lower)alkyl, the phenyl moiety of which may have halogen or nitro or amino and the alkyl moiety of which may have carboxy or esterified carboxy; halo-phenyl; morpholino; or morpholino(lower)alkyl, R R 2 and R3are each lower alkyl, X is or and Y i.-N or -NCH 2 and pharmaceutically acceptable salt thereof. Pharmaceutical compositions and methods for the treatment of diseases involving said compounds.